Int. J. Environ. Res.

Public Health 2004, 1(2), 106110

International Journal of
Environmental Research and Public Health
ISSN 1660-4601
www.ijerph.org
2004 by MDPI

Investigation into the Nephrotoxicity of Nigerian Bonny Light Crude Oil

in Albino Rats
O. E. Orisakwe1*, A. A. Njan2, O. J. Afonne1, Akumka D. D2, V. N. Orish1, O. O. Udemezue3
1
Toxicology Unit, Department of Pharmacology, College of Health Sciences, Nnamdi Azikiwe University, Nnewi Campus,
P.M.B 5001, Nnewi, Anambra State, Nigeria.
2
Department of Pharmacology and Toxicology, National Institute for Pharmaceutical Research and Development, Idu,
Abuja, Nigeria,
3
Department of Anatomy, College of Health Sciences, Abia State University, Uturu, Nigeria
*Correspondence to Dr. Orish Ebere Orisakwe. E-mail: eorish@aol.com

Received: 21 December 2003 / Accepted: 18 May 2004 / Published: 30 September 2004

Abstract: The effect of bonny-light crude oil was assessed in adult albino rats. The rats were administered
with 200, 400, and 800mg/kg body weight of the crude oil orally for 7 days. Fluid intake was measured daily,
initial and final animal body was recorded. The toxic effects on the kidneys were assessed and histological
studies carried out. The results revealed that the kidney cells were damaged; crude oil caused a destruction of
the renal reserve capacity. There was a significant increase (p 0.05) in creatinine in the high dose group
(800mg/kg), and a significant decrease (p 0.05) in urea concentration. Histological examination indicates
that crude oil induced severe pathologic changes in the forms of necrosis and oedema.

Key words: bonny light, crude oil, nephrotoxicity

Introduction introduced into the environment. There is a concern that

workers and other individuals exposed to crude oil might
Crude oil has been described as a complex mixture of have an increased incidence of organ damage. After
over 6000 potentially different hydrocarbons and metals absorption via pulmonary or gastrointestinal routes,
[1]. Crude and refined petroleum, and oil field chemicals crude oil is transported in plasma initially bound to
and emissions are highly complex chemical mixtures. albumin and other larger proteins to the liver.
Crude petroleum contains hundreds of compounds and In Nigeria, crude oil is predominantly found in the
the chemical composition varies between geologic riverine areas. Over the years the local population has
formations [2]. used crude oil for various ailments such as
Knowledge of human responses to acute exposures to gastrointestinal disorders, burns, foot rot and leg ulcers,
petroleum components comes from studies with several poisoning and witchcraft. The present study, investigates
solvents containing benzene and petroleum [3]. the renal effect of bonny-light crude oil.
Recognized human biochemical and physiological
responses associated with acute exposures to natural Materials and Methods
crudes are mainly transient and short lived unless the
concentrations of the components are unusually high Matured albino rats weighing between 124-130g
[4]). Pollution implies that a particular level of were obtained from the Animal Facility Center of the
contamination has deleterious effects, which may take Department of Pharmacology and Toxicology, National
the form of hazards to human health, interference with Institute for Pharmaceutical Research and Development,
human activities, reduction of human amenities or Abuja, Nigeria. The animals were acclimated to housing
harmful effect on living resources. Substantial amounts conditions for at least one week prior to commencement
of those potentially toxic substances have been of the experiment. Animals were housed singly at

2004 MDPI. All rights reserved.

Int. J. Environ. Res. Public Health 2004, 1 107

ambient temperature of 23 30 C and a 12 h light, 12 h the high dose group (800mg/kg) compared to the control.
dark cycle. Water and food were provided ad libitum for There were no significant differences (p 0.05) in the
the animals. absolute and relative kidney weights of the animals in
Animals were divided into 4 groups of 5 male rats the three experimental groups.
each. The three test groups received by gavage for seven The results show a significant increase (p < 0.05) in
days freshly prepared Bonny light crude oil at doses of serum creatinine in the 800mg/kg crude oil group;
200 mg/kg, 400 mg/kg and 800 mg/kg while the 4th whereas there was significant increase in potasium in the
group received a solution of Tween 80 in water only as 200 and 400mg/kg crude oil treated groups. There was
control. These doses were based on that used by the local also a significant decrease (p < 0.05) in serum urea,
population in folkloric medicine. Food and fluid intake sodium, bicarbonate and chloride in the bonny-light
were measured daily. The final body weights of the rats treated group when compared with the control (Table 2).
were recorded. The histological changes of kidney in the different
Nigerian bonny-light crude oil was obtained from treatment groups are shown in Figures 1- 4. The control
Nigerian National Petroleum Corporation (N.N.P.C.) group showed normal histological structure. Low dose
Research Laboratory Port Harcourt, Nigeria. The bonny crude oil produced slightly congested glomeruli and
light crude oil was dissolved in Tween 80. hyperemia, a congestion caused intraglomerular space
At the end of the seven days exposure period, the between the tuft and capsule. The glomeruli appeared
animals were weighed and sacrificed under chloroform infiltrated and congested creating wider spaces between
anesthesia, and serum obtained. Sodium and potassium the tuft and the capsule in the medium dose crude oil
in serum were determined with Corning 410C clinical group. Also in this group some of the glomeruli appeared
flame photometer (Corning Instruments, England) at 589 degenerated. The high dose group showed empty and
and 768nm respectively. Titrimetric method of Schales widened capsules with severely degenerated glomeruli
and Schales (1941) [5] was used for the determination of and noticeable necrosis.
chloride. The potassium and sodium were estimated
using the flame emission spectrophotometer (corning Discussion
brand). Urea was analysed spectrophotometrically at
525nm [6]. Creatinine was determined using the method Exposure of humans and animal to crude oil, which is
of Taussky 1956 [7] at 520 nm. Serum potassium, increasing in terms of the environmental levels, and
sodium, bicarbonate and chloride were measured by the application to body, may be toxic. Crude oil is used in
use of commercial kits (Biosystem, Spain). folkloric medicine in the Niger-delta area of Nigeria for
The animals were then sacrificed under chloroform the treatment of various ailments including stomach up-
anaesthesia and the kidneys were harvested, weighed and set, wound, and burns [8]. The route of administration is
fixed in 10% buffered formalin for 48 hrs. The kidneys mostly oral and external application for burns and
were processed using an automatic tissue processor, wounds. In several organs, mainly heart and liver, cell
embedded in paraffin wax, and sections (5m thick) cut damage is followed by increased levels of a number of
using a rotary microtome. The sections were stained by cytoplasmic enzymes in the blood, a phenomenon that
haematoxylin and eosin (H&E) method, and examined provides the basis for clinical diagnosis of heart and liver
and photographed by using a light microscope. Two diseases e.g. liver enzymes are usually raised in acute
Histopathologists examined the sections, independently. hepatotoxicity but tend to decrease with prolonged
intoxication due to damage to the liver cells. The
Statistical Analysis Nigerian Bonny crude oils are classified as light crude
oils, with aromatic hydrocarbons accounting for up to
The values were reported as means SEMs. Data 45% of the total hydrocarbons. As aromatic
were analyzed using the Students t-test, while Duncans hydrocarbons are relatively soluble in water [9], it is
multiple-range test was used to test for differences expected that the potential of this light crude oil to have
between treatment groups using Sigma-stat 2.0 software. adverse toxic effects is higher than for heavier, less
P < 0.05 was considered statistically significant. watersoluble crude oils. It is known that lipophilic
xenobiotics may have the characteristics of both electron
Results uncouplers and energy inhibitors [10]. Since a large
proportion of the crude oil components is lipophilic in
Table 1 shows the effect of Bonny light crude oil on nature biological membrane may be the target sites
body weight, fluid intake, absolute and relative weights where adverse effects occur.
of kidney after 7 days of sub-chronic exposure to male The kidney can suffer considerable damage before
rats. There was no significant difference in the mean losing sufficient function to modify the normal clinical
fluid intake between the three dose levels. The animals indication of renal disease such as the serum creatinine
demonstrated a progressive increase in body weight in concentration [11]. Approximately 50% or more of renal
the control and lower doses during the exposure period Table 1. Effect of Bonny light crude oil on body
and there was a significant reduction in body weight in relative weights of kidney after 7 days of sub-chronic
108 Int. J. Environ. Res. Public Health 2004, 1

Treatment Final body Absolute kidney Relative kidney Fluid intake

Dose (mg/kg) weight (g) weight (g) weight (%) ml/day/rat

Control 180.3 12.9 1.2 0.7 0.67 0.3 20.03 2.39

(11311.2)

200 166.1 7.2* 1.5 0.5 0.90 0.1 21.09 2.34

(112.6 12.9)

400 156.7 11.8* 1.3 0.7 0.83 0.2 22.89 1.18

(114.0 8.6)

800 136.8 3.6* 1.4 0.4 1.02 0.2 22.35 4.16

(124.7 21.8)
Numbers in parenthesis indicate initial body weight of rats.
N=5
*Significantly different from control at p < 0.05
Relative weight = Absolute weight x 100
Final body weight

Table 2. Effect of Bonny light crude oil on serum electrolytes, urea and creatinine after 7 days of sub-chronic
exposure to male rats.

Treatment
Dose (mg/kg) Urea Creatinine HCO3-
Cl- (mmol/l) Na+ (mmol/l) K+ (mol/l)
(mmol/l) (mmol/l) (mmol/l)

Control 7.6 1.6 117.3 2.5 26.3 3.2 97.0 2.4 145.3 6.1 5.7 2.4

200 5.7 0.2 107.0 1.7* 30.6 3.7# 94.0 2.0* 144.1 0.1 7.4 0.1#

400 4.9 0.8* 112.0 5.1* 23.0 0.2 95.0 2.0* 143.6 3.2 8.1 0.7#

800 2.4 0.1* 134.0 4.0# 21.0 0.1* 90.0 2.1* 140.6 1.5* 4.8 0.4

N = 5.
*Significantly reduced (p < 0.05) compared to control.
#
Significantly increased (p < 0.05) compared to control.
Int. J. Environ. Res. Public Health 2004, 1 109

A: Control showing normal kidney B: Low dose or 200mg/kg crude oil showing slightly
congested glomeruli

C: Medium dose or 400mg/kg crude oil showing some D: High dose or 800mg/kg crude oil showing severely
degenerated glomeruli degenerated glomeruli

Figure 1. Photomicrographs of H&E stained sections of rat kidney (Magnification X250).

capacity can be lost before serum creatinine become nephrotoxicants, with nephrotoxicity defined as
abnormal and disease is detectable clinically. A battery increased kidney weight (absolute or relative) [12].
made up of a combination of different types of test can There were no significant (p < 0.05) differences in
aid in the detection of damage by a nephrotoxin and also the absolute and relative kidney weights of the animals
allows for the determination of various threshold in the three test groups. This observation seems not to
damage. The detection of renal damage at a reversible conform with the above definition of toxicity probably
stage is necessary before effective preventive measures due to the short duration of the exposure to the bonny-
can be taken to halt the progress of damage to the light crude oil. However the significant (p < 0.05)
irreversible stage. increase in serum creatinine and potasium in the treated
The absolute and relative kidney weights, as well as groups and also the significant (p < 0.05) decrease in
body weight gain were significantly (p < 0.05) decreased serum urea, sodium, bicarbonate and chloride in the
by exposure to the bonnylight crude oil. Researchers bonny-light treated group when compared with the
have reported that absolute kidney weight is a relatively control is suggestive of renal pathology. This in part is in
sensitive indicator of nephrotoxicity for known agreement with Kluwe definition of renal toxicity [12].
110 Int. J. Environ. Res. Public Health 2004, 1

References 8 Orisakwe, O. E.; Akumka D. D.; Afonne O. J.;

Gamaniel K. S.: Investigation into the
1 Edwards, C. W.: Toxicology of oil field waste pharmacological basis for folkloric use of Bonny
hazards to livestock associated with the petroleum light crude oil in Nigeria. Ind. J. Pharmac., 2000,
industry. Vet. Clin. N. Amer. 1989, 5, 363-374. 32, 231-34.
2 Coppock, R. W.; Mostrom, M. S.; Khan A. Semalula 9 Volkman, J. K.; Miller, G. J.; Revill, A. T.; Connell
S. S.: Toxicology of oil field pollutants in cattle: A D. W.: Oil spills. In: Environmental Implications of
Review. Vet. Hum. Toxicol. 1995, 37 (6), 369-576. offshore oil and Gas Development in Australia - The
3 Zedeck, M. S.: Polycycic aromatic hydrocarbon a findings of an Independent Scientific Review (Swan
review. J Environ Pathol Toxicol 1980, 357-367.
J. M., Neff J. M., and Young P. C., Eds) Australian
4 Campbell, D.; Cox, D.; Crum, J.; Foster, K.;
Petroleum Exploration Association, Sydney. 1994,
Christie, P.; Brewster, D.: Initial effects of the
pp.509-695.
grounding of the tanker Braer on health in Shetland.
BMJ, 1993, 307, 251-55. 10 Masana. O.; Takaaki, M.; Fumio, I.; Kohei, E.;
5 Schales, O.; Schales S. S.: A simple and accurate Ritsue, S.; Tadamichi, M.; Bunji, I.: Classification
method for the determination of chloride in of potentially toxic chemicals based on their effects
biological fluids. J. Biol.Chem.; 1941, 140, 879-84. on mitochondrial respiration. Physiol Chem Phys
6 Coulombe, J. S.; Faureau, L. A.: A simple Med NMR, 1983, 15, 229-234.
colorimetric method for the determination of urea in 11 Longman-Adman, N.: Renal effects of
the presence of diacetylmonoxime and Environmental and occupational lead exposure.
thiosemicarbazide. Clin. Chem., 1963, 9, 102 Environ. Health.Perspect, 1997, 105, 9,938.
7 Taussky, H. H.: Modified alkaline picrate method 12 Kluwe, W. M.: Renal function tests as indicators of
determination of creatinine in serum. Clin. Chimca. kidney subacute toxicity studies. Toxicol. Appl.
Acta; 1956, 1, 200-02. Pharmacol., 1981, 57, 414-424.